Fig. 6: JHU083 treatment drives metabolic reprogramming in Mtb-infected lungs.

As described in Fig. 2a, Mtb-infected 129S2 mice were treated with JHU083, RIF, or PBS every day starting at day 1 post-infection. Mice were sacrificed at week 2, the lungs were harvested, and total metabolites were methanol (MeOH) extracted as described in the “Methods”. The total metabolites were normalized to the tissue weight and then to the untreated controls. We detected significant changes in the levels of (a) citrulline in whole lung MeOH extract (n = 5/group), (b) nitric oxide (NO) in Mtb-infected BMDMs treated with JHU083 as measured by the Griess colorimetric assay (n = 5/group), (c) 5-hydroxyindole acetic acid (5-HIAA) in whole lung MeOH extract (n = 5/group), and (d) quinolinic acid in whole lung MeOH extract (n = 5/group). Schematic representation of the (e) arginine and (f) tryptophan metabolic pathways with metabolites indicated in black and enzymes in blue. Green/red arrows next to a metabolite indicate statistically significant accumulation/depletion, respectively, in the JHU083-treated whole lung MeOH extract group compared to that for the PBS control. Abbreviations: arginine decarboxylase (ADC), L-arginine:glycine amidinotransferase (AGAT), arginase (ARG), guanidinoacetate N-methyltransferase (GAMT), nitric oxide synthase (NOS), ornithine decarboxylase (ODC), ornithine decarboxylase (OTC), dopa decarboxylase (DDC), 3-hydroxyanthranilate 3,4-dioxygenase (HAAO); indoleamine 2,3-dioxygenase (IDO), kynurenine 3-monooxygenase (KMO), kynureninase (KYNU), monoamine oxidase A/B (MAOA/B), tryptophan 2,3-dioxygenase (TDO2), tryptophan hydroxylase (TPH1/2). Data were plotted as mean ± SEM. Statistical significance was calculated using a two-tailed student t-test considering unequal distribution. The exact p-values are provided in the Source Data file. * < 0.05, ** < 0.01. NS stands for non-significant change, p-value was >0.05. The experiment was performed twice.